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DnaB, the DNA helicase, gets the energy to unwind DNA during replication by hydrolyzing ATP. When its ATPase activity is blocked, unwinding stops too. This makes the ATPase reaction a useful screen for helicase inhibitors in drug discovery.
Pack Sizes
Both sizes include the H. influenzae DNA helicase. The only difference is the number of assays: DNAB100KH provides reagents for 100 assays and DNAB500KH for 500 assays (384-well format), with component volumes scaled accordingly.
| 100 assays (DNAB100KH) | 100 assays; enzyme included |
|---|---|
| 500 assays (DNAB500KH) | 500 assays; enzyme included |
Kit Components
| Component | 100 assays (DNAB100KH) | 500 assays (DNAB500KH) |
|---|---|---|
| 10 x Assay buffer | 500 µl | 2 ml |
| 100 x DNA | 35 µl | 170 µl |
| 100 x H. influenzae DNA helicase | 30 µl | 150 µl |
| 100 x ATP | 35 µl | 170 µl |
| Phosphate detection dye (also sold separately as MicroMolar Phosphate Assay Reagent, BHT16300173) | 5 ml | 25 ml |
Final reaction conditions: 20 mM HEPES pH 7.5, 20 mM potassium glutamate, 1 mM DTT, 0.005% Triton X-100, 10 mM MgCl2, 20 µg/ml DNA, 0.25 mM ATP and 200 nM DNA helicase.
Assay Principle
The kit detects the phosphate produced when H. influenzae DNA helicase hydrolyzes ATP in the presence of DNA. A phosphate detection dye is added and absorbance is read at 650 nm; inhibitors of the helicase ATPase lower the signal.
Specifications
| Assay Target | H. influenzae DNA helicase ATPase |
|---|---|
| Target Type | Helicase |
| Organism | H. influenzae |
| Assay Type | Enzyme Activity Assay; Inhibitor Screening Assay |
| Detection Method | Colorimetric |
| Detection Wavelength | Absorbance 650 nm |
| Plate Format | 384-well plate |
| Instrument Required | Absorbance microplate reader |
| Assay Time | About 65 min (60 min reaction + 5 min detection) |
| Sizes | 100 assays, 500 assays |
| Storage | Please contact us to confirm storage conditions. |
| Shipping | Shipped frozen. |
Protocol
The manufacturer's protocol describes the procedure and reaction volumes.
- H. influenzae DNA Helicase ATPase assay (PDF, see Documents): Covers catalog # DNAB100KH, DNAB500KH.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this H. influenzae DNA helicase ATPase assay — including larger pack sizes, bulk reagents, bulk enzyme, or a protocol adapted to your samples. Click Talk to a Scientist, email support@biohippo.com, or explore our Research Services. Our team will be in touch shortly.